Development of a highly sensitive assay for enzyme‐mediated reductive degradation of polychlorinated dibenzo‐p‐dioxin
Development of a highly sensitive assay for enzyme‐mediated reductive degradation of polychlorinated dibenzo‐p‐dioxin
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DOI：
10.1002/etc.1775
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发表时间：
2012-05
影响因子：
4.1
通讯作者：
Yuzo Suzuki;M. Nakamura;Y. Otsuka;Nao Suzuki;Keisuke Ohyama;T. Kawakami;Kanna Sato;S. Kajita;S. Hishiyama;A. Takahashi;Y. Katayama
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文献类型：
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作者：
Yuzo Suzuki;M. Nakamura;Y. Otsuka;Nao Suzuki;Keisuke Ohyama;T. Kawakami;Kanna Sato;S. Kajita;S. Hishiyama;A. Takahashi;Y. Katayama
监测了2-氯-4，5-O-（4′-甲基-7 ′，8′-二苯基）醚（CMDPE）（2，7-二氯二苯并对二恶英（2，7-DCDD）的类似物）在Geobacillus sp. UZO 3无细胞提取物介导下的降解。在65°C下孵育18小时的完全反应混合物的乙酸乙酯提取物通过薄层色谱（TLC）分级分离结合光谱检测或通过气相色谱-质谱（GC-MS）分析。通过TLC成功分离反应产物4-甲基伞形酮（4 MU），并通过透射仪在450 nm处观察。通过GC-MS成功检测到4 MU、4-氯苯酚和反应中间体6-氯苯氧基-4-甲基伞形酮。这些化合物的存在表明，Geobacillus sp. UZO 3无细胞提取物也以先前在2，7-DCDD中报道的类似机制催化CMDPE的二芳基醚键的还原裂解。在本研究中，作者描述了一种简单而高灵敏度的荧光检测新的二恶英降解酶（S）。Environ.毒理学. 2012; 31：1072-1075。© 2012 SETAC
The degradation of 2‐chloro‐4,5‐O‐(4′‐methyl‐7′, 8′‐diphenyl)ether (CMDPE), an analog of 2,7‐dichlorodibenzo‐p‐dioxin (2,7‐DCDD), mediated by Geobacillus sp. UZO 3 cell‐free extract was monitored. Ethyl acetate extracts of a complete reaction mixture incubated at 65°C for 18 h were analyzed either by thin layer chromatography (TLC) fractionation coupled with spectrometric detection or by gas chromatography–mass spectrometry (GC‐MS). The reaction product 4‐methylumbelliferone (4MU) was successfully isolated by TLC and visualized by a transilluminator at 450 nm. The 4MU, 4‐chlorophenol, and reaction intermediate 6‐chlorophenoxy‐4‐methylumbelliferone were all successfully detected by GC‐MS. The presence of these compounds suggest that Geobacillus sp. UZO 3 cell‐free extract also catalyzes the reductive cleavage of the diaryl ether bonds of CMDPE in a similar mechanism previously reported in 2,7‐DCDD. In the present study, the authors describe a simple and highly sensitive fluorescent assay for a new dioxin degrading enzyme(s). Environ. Toxicol. Chem. 2012; 31: 1072–1075. © 2012 SETAC